Device for performing biopsies on a vesicular object

ABSTRACT

A device for removing material from a vesicular object, said device being a pipette having a sealed tip at its distal end and an aperture whereby the distal tip of the device is inserted into the target object, the aperture aligned with the material to be removed from the object, vacuum applied to the inside of the device, such vacuum drawing the material into the aperture, and the device being removed from the object in such a manner as to cut or otherwise separate the material in the pipette from the object, the material remaining in the pipette, while leaving the object undamaged.

CROSS-REFERENCE TO RELATED APPLICATIONS

1. This application is a division of application Ser. No. 13/986,029filed Mar. 25, 2013, now U.S. Pat. No. ______ [fill in later], granted______ [fill in later].

2. This application also claims the benefit of PPA Ser. No. 61/686,025filed Mar. 28, 2012 by the present inventor, which is incorporated byreference.

FEDERALLY SPONSORED RESEARCH

This invention was not made under a government contract and thegovernment has no rights in it.

SEQUENCE LISTING OR PROGRAM

Not applicable.

BACKGROUND Prior Art

The following is a tabulation of some prior art that presently appearsrelevant:

U.S. PATENTS

PAT. NO. ISSUE DATE PATENTEE 8,394,033 July 2006 DiCarlo 8,376,957February 2013 Hibner 20060200040 September 2006 Weikel 8,357,103 January2013 Mark

BACKGROUND

The present method and device relates to a method and device forremoving tissue or other cellular material (cellular material) from avesicular object having a size typically in the neighborhood of 100-300microns. This device has particular application for removing cellularmaterial from mammalian embryos at the hatched blastocyst stage ofdevelopment. This has not been possible with previous devices which havebeen mostly designed for biopsies of a much larger nature.

Removing material from the hatched blastocyst has been difficult becausethe blastocysts are spherical vesicular structures consisting of a thindouble layer of living cells surrounding a relatively large centralcavity that is filled with an aqueous fluid. The difficulty, and thefailure of the prior art, arises because of the physical characteristicsof the envelope of these living cells which is flimsy but resistant topuncture. For example, attempts to puncture a hatched blastocyst oftensimply compress the envelope of the embryo without puncturing it. Withthe present device there is no compression at all.

ADVANTAGES OF THE EMBODIMENT

The present device overcomes the above-described difficulties byproviding a device wherein the object is firmly held, for example, byusing a vacuum as described in application Ser. No. 13/374,195, theobject is then pierced by the probe. An aperture, or biopsy port, on oneside of the probe is then aligned with the material to be removed. Avacuum applied to the interior of the probe draws the cellular materialinto the probe itself for subsequent removal from the vesicular object.

An additional advantage lies in fact that the device leaves the objectunharmed after the biopsy. As an example only, the puncture wound madein the surface of an expanding or hatched blastocyst by the biopsy probewill involve a very small percentage (less than 1%) of the surface. In aliving embryo (blastocyst), this wound is surrounded by rapidly dividingcells on all sides. As soon as the probe is withdrawn, the wound closestightly enough to prevent leakage of fluid into or out of the blastocoelspace and will heal closed completely within a few hours.

A yet additional advantage is that smaller biopsy probes, down to 4 umoutside diameter (OD) near the tip end, with biopsy ports at least assmall as 2 um diameter, can be made at present. This is small enough tobe used on certain large individual cells, resulting in the potential ofremoving or inserting cellular components. Smaller probes and biopsyports are possible.

SUMMARY, CONCLUSION, RAMIFICATIONS, AND SCOPE

The above-described device punctures and allows removal of cellularmaterial from a very specific location on a vesicular object. Forexample, an identifiable portion of the surface of a hatched blastocystis the Inner Cell Mass, consisting of embryonic stem cells. This devicehas the potential to harvest embryonic stem cells without serious damageto the embryo.

BRIEF DESCRIPTION OF THE DRAWINGS

In order that the device may be clearly understood and readily carriedinto effect, a preferred embodiment of the invention will now bedescribed, by way of example only, with reference to the accompanyingdrawings wherein:

FIG. 1 is a drawing of a probe 20 microns in diameter at the location ofthe aperture, which is specified between 9 and 15 microns diameter.

FIG. 2, is a drawing representing cellular material from the object'senvelope of cells being drawn into the biopsy probe through the port.

REFERENCE NUMERALS FIG. 1.

-   1. Aperture.-   2. Sealed distal tip.-   3. Pipette's hollow center.

FIG. 2.

-   4. Envelope of cells.-   5. Blastocoel fluid.-   6. Biopsy sample.

DETAILED DESCRIPTION OF AN EMBODIMENT Elements

A device for performing biopsies on a vesicular object is shown inFIGS. 1. and 2. In this embodiment the device includes a hollow pipettewith a sealed tip at its distal end. Such pipette having an aperture (1)through the pipette wall (biopsy port) located only a few microns,typically 40 to 70 um, proximally from the sealed pipette's distal tip.(2). More specifically, the biopsy port is located on one sidewall ofthe pipette, at a right angle to the pipette's long axis, communicatingwith the pipette's hollow center and is created, as explained in moredetail below, without damage to the pipette's far wall.

In the present embodiment, the diameter of the biopsy port isapproximately one-half of the outside diameter of the pipette and theport is created in such a way as to leave the edges of the port sharpand capable of cutting or shearing cellular material from a target.

Such pipette, with the sealed distal tip and biopsy port, being referredto herein as a biopsy probe.

Method of Manufacture

A standard method of manufacture for the biopsy probe is first pulling astandard or thin-wall 1.5 mm OD (outside diameter) glass tube to a longtaper, then scoring and breaking the thin glass tip at the desireddiameter. This diameter has been, to date, typically between Bum and 25um OD. Then, on a microforge at relatively low temperature, just hotenough to melt the glass, the tip end is touched to the molten glassbead, which seals it and draws the molten tip into a sharp spike.

The aperture, or biopsy port, on the formed probe is then drilled fromthe side using an appropriate device, for example only, a femtosecondlaser. The laser must be tuned to make a clean hole of the desireddiameter through only the near sidewall of the glass tube, withoutdamage to the inside of the far sidewall. The position of the port isdetermined by the diameter of the probe near the tip. For example only,if the diameter is “x”, the port diameter is “0.5x” and the port ispositioned “5x” to “7x” back from the sharp, and sealed, distal tip.This allows the biopsy port to be positioned (for use on an early embryofor example) at the level of the envelope of living cells when the sharptip end is still inside the blastocoel space. The intended use of theprobe, e.g. type and stage of development of an embryo to be biopsiedand physical size of the cells to be captured, determines the specificposition and size of the biopsy port.

Operation

A standard operation of the probe would include the following steps. Thetarget is captured and held and the sharp and sealed distal tip of thebiopsy probe is advanced, possibly as described in application Ser. No.13/374,195. After penetrating a specific area of the object's surface,the biopsy probe's sealed distal tip (2) is advanced into the centralspace of the target (5) until the biopsy port (1) is aligned with thehole in the surface of the target made by the biopsy probe's sealeddistal tip, or aligned in close proximity to the material to be removedfrom the interior of the object. A vacuum is then applied inside thebiopsy probe and material is drawn into the biopsy probe through theport. (6) As the biopsy probe is withdrawn from the object, if thematerial is connected to the object, the material in the biopsy port issheared, cut, or torn, away from the object leaving the objectundamaged.

While the above description contains many specificities, these shouldnot be construed as limitations on the scope of any embodiment, but asexamples of various embodiments thereof. Many other ramifications andvariations are possible within the teachings of the various embodimentswithout departing from the scope of the method and device disclosedabove.

Thus the scope should be determined by the appended claims and theirlegal equivalents, and not by the examples given.

I claim:
 1. A device for removing material from a vesicular object(object) said device being a pipette and having: a, a sealed tip at itsdistal end (2), the distal tip being sharp enough to puncture theobject; b, an aperture (1), both the specific location and size of theaperture being appropriate for the object and the material to beremoved; and c, the aperture having edges sharp enough to cut orotherwise separate the material in the pipette from the object; wherebythe distal tip of the device is inserted into the object, the aperturealigned with the material to be removed from the object, vacuum appliedto the inside of the device, such vacuum drawing the material into theaperture, and the device being removed from the object in such a manneras to cut or otherwise separate the material in the pipette from theobject, this material remaining in the pipette, while leaving the objectundamaged.